Influence of bone adaptation on tendon‐to‐bone healing in bone tunnel after anterior cruciate ligament reconstruction in a rabbit model

Anterior cruciate ligament (ACL) reconstruction with placement of grafted tendon in bone tunnel is a common surgical procedure. Bone tunnel creation may result in stress shielding of postero‐lateral regions of tibial tunnel. The present study was designed to characterize the changes of peri‐graft bone and compare with tendon‐to‐bone (T‐B) healing in spatial and temporal manners after ACL reconstruction in rabbit. Surgical reconstruction using digital extensor tendon in bone tunnel was performed on 48 rabbits. Twelve rabbits were sacrificed at 0, 2, 6, and 12 weeks postoperatively for radiological and histological examinations. Bone mass and microarchitecture at the anterior, posterior, medial, and lateral regions of tunnel wall at distal femur and proximal tibia were evaluated. Using peripheral quantitative computed tomography, a 26, 22, and 42% decrease in bone mineral density (BMD) relative to baseline was present in the medial region of the femoral tunnel and the posterior and lateral regions of the tibial tunnel, respectively, at week 12 postoperatively (p < 0.05). It was accompanied by a decrease in trabecular number and increase in trabecular spacing, the shift of platelike to rodlike trabeculae, and loss of anisotropy under micro‐computed tomography evaluation. This finding was echoed by histology showing increased osteoclastic activities and poor T‐B healing in these regions. In conclusion, the postoperative bone loss and associated poor T‐B healing was region‐dependent, which may result from adaptive changes after tunnel creation. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1447–1456, 2009     

生物骨传导性骨水泥促进前交叉韧带重建术腱骨愈合的研究进展(英文)

近10年来,应用软组织移植物行前交叉韧带重建术越来越普遍。手术的远期疗效主要取决于肌腱移植物能否在骨隧道内达到坚强的腱-骨愈合。但是,目前面临的问题是肌腱移植物在骨隧道内获得腱-骨愈合所需要的时间相当长。研究发现,在腱-骨界面局部应用生物骨传导性的骨水泥能有效地促进肌腱移植物的骨愈合。这类骨水泥主要是磷酸钙。本文就生物骨传导性骨水泥促进前交叉韧带重建术腱-骨愈合的研究进展作一综述。     

Alendronate prevents bone loss and improves tendon-to-bone repair strength in a canine model.

Previously we showed a loss of bone and a concomitant decrease in mechanical properties in the first 21 days after flexor tendon insertion site injury and repair in a canine model. The goal of this short-term study was to suppress bone loss after insertion site repair using alendronate in an attempt to prevent the reduction in biomechanical properties. Flexor tendons of the second and fifth digits of the right forelimbs of canines were injured and repaired. Dogs received a daily oral dose of alendronate (2 mg/kg). One digit in each dog also received a local dose of alendronate in the bone tunnel at the time of surgery. The repair was evaluated for bone mineral density (BMD) and biomechanical properties and compared to data from a previous study in which no alendronate was used. Alendronate was effective in protecting the distal phalanx from resorption during tendon-to-bone healing (BMD was 94 and 104% of control for systemic alendronate and for systemic plus local alendronate, respectively). Alendronate treatment prevented much of the decrease in ultimate load that occurs in the first 21 days. Without treatment, ultimate load was 42% of control. With systemic alendronate treatment and systemic plus local alendronate treatment, ultimate load was 78 and 69% of control, respectively. Failure mode was significantly different when comparing alendronate treatment to repair alone. A lower incidence of suture pull through was found in alendronate treated dogs, suggesting less tendon degeneration. Ultimate load can be improved in association with preventing the bone loss that normally occurs during the early period following tendon-to-bone repair. These initial short-term data demonstrate the potential for a clinical treatment that could enhance tendon-to-bone healing.     

Comparison of volumetric bone mineral density in the operated and contralateral knee after anterior cruciate ligament and reconstruction: A 1‐year follow‐up study using peripheral quantitative computed tomography

The purpose of this study was to quantify changes in volumetric bone mineral density (vBMD) in the tibial plateau of the operated and contralateral leg measured using peripheral quantitative computed tomography (pQCT) before and 3, 6, and 12 months after anterior cruciate ligament (ACL) reconstruction. The ACL was reconstructed with a hamstring tendon autograft using press‐fit fixation. pQCT measurements of the proximal tibia were obtained in 61 patients after ACL reconstruction, and total, cortical, and trabecular vBMD were calculated. vBMD in the operated leg decreased from baseline to 3 months (?12% [total], ?11% [cortical], and ?12.6% [trabecular]; p < 0.001) and remained below baseline for 12 months after surgery (6 months: ?9.5%, ?9.4%, and ?9.6%, p < 0.001; 12 months: ?8%, ?5%, and ?11%, p < 0.001). vBMD in the contralateral leg was slightly reduced only 6 months after surgery. Including age and sex as covariates into the analysis did not affect the results. ACL reconstruction contributed to loss in bone mineral density within the first year after surgery. The role of factors such as time of weight‐bearing, joint mechanics, post‐traumatic inflammatory reactions, or genetic predisposition in modulating the development of posttraumatic knee osteoarthritis after ACL injury should be further elucidated. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1804–1810, 2015.

     

Differences in Tendon Graft Healing Between the Intra-articular and Extra-articular Ends of a Bone Tunnel

The basic biology of healing between a tendon graft and bone tunnel remains incompletely understood. Distinct variability in the morphological characteristics of the healing tendon–bone attachment site has been reported. We hypothesized that spatial and temporal differences in tendon-to-bone healing exist at different regions of a surgically created bone tunnel. Twenty-four male, Sprague–Dawley rats underwent anterior cruciate ligament (ACL) reconstruction in the left knee using a flexor digitorum longus tendon graft secured using suspensory periosteal fixation. Animals were sacrificed at 4, 7, 11, 14, 21, and 28 days after surgery and prepared for routine histology and immunohistochemical analysis of the healing enthesis at the intra-articular aperture (IAA), mid-tunnel, and extra-articular aperture (EAA). Six animals were used to measure mineral apposition rate (MAR) along the healing bone tunnel by double fluorochrome labeling at 14 and 28 days after surgery. The total area of calcified bone matrix was assessed with von Kossa staining and Goldner-Masson trichrome staining, respectively. The healing tendon–bone interface tissue exhibited a wide chondroid matrix at the IAA, in contrast to a narrow, fibrous matrix at the EAA. There were significantly more osteoclasts at the IAA compared to EAA throughout the study period, except 4 days after surgery (p < 0.05). Collagen continuity between the tendon graft and bone tunnel increased over time, with a more parallel orientation and increased collagen fiber continuity between tendon and bone at the EAA compared to the IAA. MAR was also significantly greater at the EAA at 4 weeks (p < 0.001). Significant differences in healing between the tendon graft and bone exist along the length of bone tunnel secured with suspensory fixation. The etiology of these differences is likely multifactorial in nature, including variable biological and biomechanical environments at different ends of the tunnel. Understanding these differences may ultimately allow surgeons to improve the quality of graft fixation and long-term outcomes after ACL reconstruction.     

Local bisphosphonate treatment increases fixation of hydroxyapatite‐coated implants inserted with bone compaction

It has been shown that fixation of primary cementless joint replacement can independently be enhanced by either: (1) use of hydroxyapatite (HA) coated implants, (2) compaction of the peri‐implant bone, or (3) local application of bisphosphonate. We investigated whether the combined effect of HA coating and bone compaction can be further enhanced with the use of local bisphosphonate treatment. HA‐coated implants were bilaterally inserted into the proximal tibiae of 10 dogs. On one side local bisphosphonate was applied prior to bone compaction. Saline was used as control on the contralateral side. Implants were evaluated with histomorphometry and biomechanical push‐out test. We found that bisphosphonate increased the peri‐implant bone volume fraction (1.3‐fold), maximum shear strength (2.1‐fold), and maximum shear stiffness (2.7‐fold). No significant difference was found in bone‐to‐implant contact or total energy absorption. This study indicates that local alendronate treatment can further improve the fixation of porous‐coated implants that have also undergone HA‐surface coating and peri‐implant bone compaction. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:189–194, 2009     

Role of osteoblast–fibroblast interactions in the formation of the ligament‐to‐bone interface

The anterior cruciate ligament (ACL) inserts into bone through a characteristic fibrocartilagenous interface, which is essential for load transfer between soft and hard tissues. This multi‐tissue interface is lost post ACL reconstruction, and the lack of an anatomic fibrocartilage interface between graft and bone remains the leading cause of graft failure. Currently, the mechanism of interface formation is not known. As a fibrocartilage‐like tissue is found within the bone tunnel post ACL reconstruction, we hypothesize that fibroblast–osteoblast interactions at the graft‐to‐bone junction play a role in fibrocartilage formation. To test this hypothesis, a co‐culture model permitting osteoblast–fibroblast communications was used to determine the effects of heterotypic interactions on cell phenotype and the development of fibrocartilage‐relevant markers in vitro. It was found that co‐culture decreased cell proliferation and osteoblast‐mediated mineralization, while inducing fibroblast‐mediated mineralization. Moreover, the expression of interface‐relevant markers such as collagen type II and aggrecan were detected. Our findings suggest that osteoblast–fibroblast interactions may lead to cell trans‐differentiation and eventual fibrocartilage formation. These results provide new insight into the mechanism of fibrocartilage formation, which are critical for interface tissue engineering and achieving biological fixation of soft tissue grafts to bone. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1609–1620, 2007     

ACL reconstruction using bone‐tendon‐bone graft engineered from the semitendinosus tendon by injection of recombinant BMP‐2 in a rabbit model

We attempted to generate a bone‐tendon‐bone structure by injecting human‐type recombinant human bone morphogenetic protein‐2 (rhBMP‐2) into the semitendinosus tendon, and an anterior cruciate ligament (ACL) defect was reconstructed by grafting the engineered bone‐tendon‐bone graft. Two ossicles with a separation distance of 1 cm were generated within the left semitendinosus tendon of a rabbit 6 weeks after the injection of rhBMP‐2 (15 µg at each site). The engineered bone‐tendon‐bone graft was transplanted in order to reconstruct the ACL by passing the graft through the bone tunnels. In the control group, the ACL was reconstructed with the semitendinosus tendon without BMP‐2 using the same methods as those used in the experimental group. The animals were harvested at 4 or 8 weeks after surgery and examined by radiographic, histological, and biomechanical methods. In the experimental group, ossicles in the bone‐tendon‐bone graft were successfully integrated into the host bone of the femur and tibia. Histological analysis revealed that characteristic features identical to the normal direct insertion morphology had been restored. Biomechanical pull‐out testing showed that the ultimate failure load and stiffness of the reconstructed ACL in the experimental group were significantly higher than those in the control group at both 4 and 8 weeks (p < 0.05). These results indicate the potential of regenerative reconstruction of the ACL, and the reconstruction resulted in the restoration of morphology and function equivalent to those of the normal ACL. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29:1923–1930, 2011     

Effect of ACL graft material on joint forces during a simulated in vivo motion in the porcine knee: Examining force during the initial cycles

This study compared three‐dimensional forces in knees containing anterior cruciate ligament (ACL) graft materials versus the native porcine ACL. A six‐degree‐of‐freedom (DOF) robot simulated gait while recording the joint forces and moments. Knees were subjected to 10 cycles of simulated gait in intact, ACL‐deficient, and ACL‐reconstructed knee states to examine time zero biomechanical performance. Reconstruction was performed using bone‐patellar tendon‐bone allograft (BPTB), reconstructive porcine tissue matrix (RTM), and an RTM‐polymer hybrid (Hybrid). Forces and moments were examined about anatomic DOFs throughout the gait cycle and at three key points during gait: heel strike (HS), mid stance (MS), toe off (TO). Compared to native ACL, each graft restored antero‐posterior (A‐P) forces throughout gait. However, all failed to mimic normal joint forces in other DOFs. For example, each reconstructed knee showed greater compressive forces at HS and TO compared to the native ACL knee. Overall, the Hybrid graft restored more of the native ACL forces following reconstruction than did BPTB, while RTM grafts were the least successful. If early onset osteoarthritis is in part caused by altered knee kinematics, then understanding how reconstruction materials restore critical force generation during gait is an essential step in improving a patient's long‐term prognosis. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1458–1463, 2014.     

Indian hedgehog signaling and the role of graft tension in tendon‐to‐bone healing: Evaluation in a rat ACL reconstruction model

The structure and composition of the native enthesis is not recapitulated following tendon‐to‐bone repair. Indian Hedgehog (IHH) signaling has recently been shown to be important in enthesis development in a mouse model but no studies have evaluated IHH signaling in a healing model. Fourteen adult male rats underwent ACL reconstruction using a flexor tendon graft. Rats were assigned to two groups based on whether or not they received 0N or 10N of pre‐tension of the graft. Specimens were evaluated at 3 and 6 weeks post‐operatively using immunohistochemistry for three different protein markers of IHH signaling. Quantitative analysis of staining area and intensity using custom software demonstrated that IHH signaling was active in interface tissue formed at the healing tendon‐bone interface. We also found increased staining area and intensity of IHH signaling proteins at 3 weeks in animals that received a pre‐tensioned tendon graft. No significant differences were seen between the 3‐week and 6‐week time points. Our data suggests that the IHH signaling pathway is active during the tendon‐bone healing process and appears to be mechanosensitive, as pre‐tensioning of the graft at the time of surgery resulted in increased IHH signaling at three weeks. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:641–649, 2016.     

ACL Reconstruction Graft Angle and Outcomes: Transtibial vs Anteromedial Reconstruction

Background

The importance of creating an anatomic anterior cruciate ligament (ACL) reconstruction has been receiving significant attention. The best technique by which to achieve this anatomic reconstruction continues to be debated. The two most common methods are the transtibial (TT) and anteromedial (AM) techniques. Each has its advantages and disadvantages, and the literature comparing the two remains uncertain.

Questions/Purposes

In this prospective comparative study, we aimed to compare the ACL graft and tunnel angles achieved using the anatomic transtibial (TT) and anteromedial (AM) techniques; compare the ACL graft and tunnel angles in knees that have undergone ACL reconstruction and knees with intact ACLs; and determine whether differences in the graft or tunnel angle produce differences in clinical outcomes, as measured using both physical exam and patient-reported outcomes, after ACL reconstruction.

Methods

Patients who underwent primary ACL reconstruction with bone–tendon–bone grafts using a TT or AM technique were included. Femoral graft angle (FGA), tibial graft angle (TGA), and sagittal orientation of the reconstructed ACL and contralateral native ACL were measured on post-operative magnetic resonance imaging. Post-operatively, patients underwent measurement of knee stability and completed the Knee Injury and Osteoarthritis Outcome Score (KOOS) survey.

Results

Twenty-nine patients were enrolled (AM group, 14; TT group, 15); at follow-up, KOOS data were available for 26 patients (13 in each group). There were no differences in sagittal ACL graft angle between groups or in comparison with the normal knee. The FGA was more vertical after TT reconstructions; the TGA was comparable between groups. There were no significant differences in 2-year post-operative physical exam measurements or in KOOS scores.

Conclusion

Anatomic ACL angle was restored after reconstruction with both the TT and AM techniques, despite different FGAs. No significant differences in clinical outcome were noted between groups on physical exam or KOOS at 2 years after surgery. These results suggest that TT reconstruction results in a graft position similar to that seen in AM reconstruction and that the location of the intra-articular tunnel aperture matters more than the orientation of the tunnel.

     

The effects of local and systemic alendronate delivery on wear debris‐induced osteolysis in vivo

We investigated the effects of locally and systemically administered alendronate on wear debris‐induced osteolysis in vivo. Endotoxin‐free titanium particles were injected into rabbit femurs, prior to insertion of a nonweight‐bearing polymethylmethacrylate plug into the distal femur canal. Then the particles were repeatedly injected into the knee 2, 4, and 6 weeks after the implantation. Alendronate was incorporated at three different concentrations (0.1, 0.5, and 1.0 wt %) into bone cement for local delivery. For systemic delivery, alendronate was subcutaneously injected (1.0 mg/kg/week) 1 week after the implantation and then once a week until sacrifice. Eight weeks postoperatively, there was significant evidence of osteolysis surrounding the plug in the control group compared with markedly blocked osteolysis in the 0.5 wt % and the 1.0 wt % groups, and the systemic group. There was a concentration‐dependent effect of alendronate‐loaded bone cement on the improvement of peri‐prosthetic bone stock. Notably, no significant differences were found between the 0.5 wt % and the systemic group in peri‐prosthetic bone stock and implant fixation. Collectively, although the biological efficacy after the systemic delivery of alendronate was slightly higher than that in the local treatment groups, alendronate‐loaded bone cement may be therapeutically effective in inhibiting titanium particle‐induced osteolysis in vivo. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:893–899, 2010     

前十字韧带重建中的移植物位置偏移

目的 计算可吸收界面螺钉导致的移植物偏离隧道位移,探讨其对前十字韧带重建产生的影响.方法 19个新鲜尸体膝关节标本,随机选取5个,采用7 mm、8 mm、9 mm界面螺钉固定自体肌腱,测定偏移距离.另外14个膝关节分为等长组和解剖组,等长组膝关节测量界面螺钉固定后及校正位置的移植物拉长距离;解剖组膝关节于膝关节生物力学测试仪上分别测定ACL完整组、ACL缺失组、偏移组和校正组在134 N前向负荷下膝关节屈曲0°、15°、30°、60°和90°位的胫骨前向位移.结果 (1)肌腱偏移:直径7mm、8 mm、9mm的界面螺钉分别使移植物偏移(2.36±0.11)mm、(2.72±0.06)mm、(3.00±0.06)mm.(2)等长性:初始拉长小于3 mm,偏移拉长大于3 mm,校正拉长小于3 mm.(3)生物力学:屈膝0°、15°位,ACL完整组与偏移组、校正组差异无统计学意义.屈膝30°、60°、90°位ACL完整组与其他各组比较差异均有统计学意义,屈膝30°、60°位偏移组与校正组比较差异有统计学意义.结论 无论等长重建还是解剖重建,界面螺钉均影响移植物的股骨隧道口位置.前十字韧带重建预先校正股骨隧道口位置,移植物基本会处于预先的理想位置.

Abstract：

Objective To investigate the impact of graft position shift on anterior cruciate ligament reconstruction induced by femoral fixation of interference screw. Methods Nineteen fresh cadaveric knees were used and assigned to three groups. 1) Study of graft position shift: 5 knees were randomly selected, interference screws of 7 mm, 8 mm and 9 mm were used in autologous tendon fixation, then the graft position shift were measured. 2) Study of isometry: 7 knees were randomly divided into the isometric reconstruction group (D group). In the D group, Retrobutton, interference screw and interference screw in location-corrected bone tunnel were used respectively as fixation. The isometry of grafts was evaluated. 3) Study of tibia anterior translation: 7 knees were randomly divided into the anatomic reconstruction group (J group). In the J group,the tibia anterior translation was measured in four different conditions in the same joint: intact knee joint,knee joint without ACL, ACL anatomic reconstruction by interference screw fixation, and ACL anatomic reconstruction by interference screw fixation with corrected bone tunnel location. Results 1) With 7 mm, 8mm and 9 mm interference screw fixation, graft position shift were (2.36±0.11) mm, (2.72±0.06) mm and (3.00±0.06) mm respectively. 2) Graft length change: graft length change in Retrobutton group and corrected bone tunnel group were less than 3 mm, while graft length change in those fixed with interference screw were stretched in more than 3 mm. 3) Study of tibia anterior translation: there was no difference among the intact group, the anatomic group and the corrected group at 0° and 15°. However, the difference was found between the intact group and other groups at 30°、60° and 90° of flexion, as well as between these two reconstructed methods at 20° joint flexion (P＜0.05). Conclusion In both isometric and anatomic ACL reconstruction with interference screw, the graft is pushed tightly toward the femoral tunnel wall, which shifts the graft away from the desired position. In our study we find out that the corrected location of the femoral bone tunnel significantly improves the isometry of ACL reconstruction and anatomic reconstruction.     

前交叉韧带单束重建中股骨椭圆隧道技术与圆隧道技术的比较研究

目的通过与传统圆隧道技术相比,探讨前交叉韧带(anterior cruciate ligament,ACL)单束重建中采用椭圆隧道技术制备股骨隧道的疗效及优势。方法回顾分析2016年3月-2018年2月125例采用自体肌腱单束解剖重建ACL且符合选择标准的患者临床资料,其中43例术中采用椭圆隧道技术制备股骨隧道(A组),82例采用圆隧道技术(B组)。两组患者年龄、性别、体质量指数、病程、损伤侧别、损伤原因以及术前Lysholm评分、国际膝关节文献委员会(IKDC)评分、Tegner评分、KT-1000测量值等一般资料比较,差异均无统计学意义(P>0.05)。记录两组患者术后3、6、12、24个月Lysholm评分、IKDC评分和Tegner评分,采用KT-1000评价关节稳定性。术后1d三维CT评估股骨和胫骨隧道位置。术后6、12、24个月MRI检查,测量ACL移植物近、中、远端信号/噪声比(signal/noise quotient,SNQ)。对行二次关节镜检查患者评价移植物完整性、滑膜覆盖以及张力情况。结果两组患者均获随访,随访时间12~26个月,平均23个月。A组2例、B组5例发生切口红肿,B组1例发生胫骨隧道裂纹骨折,A组1例发生膝关节屈曲活动度受限。除术后3个月Tegner评分外,其余各时间点A组Lysholm、IKDC、Tegner评分均明显高于B组(P<0.05)。术后各时间点A组KT-1000测量值亦明显小于B组(P<0.05)。术后1 d三维CT检查示两组股骨及胫骨隧道均位于ACL止点印迹内。MRI复查两组均无移植物断裂及明显松弛发生。术后6个月两组移植物中、远端SNQ差异无统计学意义(P>0.05),近端A组明显低于B组(P<0.05);术后12、24个月A组移植物近、中、远端SNQ均低于B组(P<0.05)。A组21例及B组38例患者进行二次关节镜检查,两组移植物完整性、滑膜覆盖及张力比较,差异均无统计学意义(P>0.05)。结论与传统圆隧道技术相比,ACL单束重建术中采用椭圆隧道技术制备股骨隧道,术后移植物成熟度更好,患者能获得更好膝关节功能。     

后交叉韧带重建中股骨侧"锐角效应"的相关研究及骨道技术的评价

[目的]探讨后交叉韧带重建中股骨侧的"锐角效应"以及对骨道技术进行评价.[方法]应用猪的屈趾肌腱作为移植物重建后交叉韧带,分别建立与股骨髁间窝侧壁呈80°、90°、100°夹角的骨道,以及对骨道边缘进行打磨处理.在骨道边缘与移植物之间放置压敏片计算得压强值来表示磨损作用的大小,压强值越大表示磨损作用越大.通过比较应用骨道技术重建PCL组与对照组的压强值来说明这种减小"锐角效应"的方法是否可行.[结果]应用骨道技术重建后交叉韧带各组中移植物与股骨髁间窝侧壁夹角为100°时压强最小[(3.55±0.21)MPa],并且对骨道边缘进行打磨后压强值[(3.29±0.19)MPa]明显低于对照组[(3.55±0.21)MPa].[结论]应用骨道技术可以有效减小后交叉韧带重建中股骨侧的"锐角效应".     

Macrophages accumulate in the early phase of tendon–bone healing

A scar tissue interface forms rather than a normal ligament insertion site following attachment of a tendon graft to bone. The specific cell types that initiate the process of tendon-to-bone healing are unknown. We hypothesized that inflammatory cell accumulation following tendon-to-bone repair results in this scar interface. We used a rodent model to examine the temporal and spatial pattern of accumulation of hematopoietic lineage cells in the early phase of tendon-to-bone healing. Thirty-six Lewis rats underwent anterior cruciate ligament (ACL) reconstruction in the left knee using a flexor digitorum longus tendon graft. Six animals were sacrificed at 4, 7, 11, 14, 21, and 28 days after surgery. Serial sections were analyzed for proliferating cells (PCNA), recruited macrophages (ED1), resident macrophages (ED2), neutrophils, T-lymphocytes (CD3), mast cells, immature progenitor cells/pericytes (expressing the NG2 cell-surface chondroitin sulfate proteoglycan), and newly-formed blood vessels (Factor VIII). Neutrophils, ED1(+) and ED2(+) macrophages accumulated sequentially in the healing tendon graft, with progressive cell ingrowth from the interface towards the inner tendon. Neutrophils and ED1(+) cells were seen in the tendon-bone interface at 4 days after surgery, while ED2(+) macrophages were not identified until 11 days. These cells progressively repopulated the tendon graft. NG2-positive progenitor cells were found along the edge of the bone tunnel in the interface, but these cells did not invade the tendon. Occasional T-lymphocytes and mast cells were seen in the tendon-bone interface. There was no proliferation of intrinsic tendon cells, indicating that the tendon does not directly contribute to healing. We hypothesize that cytokines produced by infiltrating macrophages are likely to contribute to the formation of a fibrous scar tissue interface rather than a normal insertion site.     

Interarticular bone tunnel healing

Purpose: To evaluate the healing behavior of an interarticular bone tunnel exposed continuously to a synovial environment. Type of Study: Experimental in vivo animal model. Methods: Twenty-six adult rabbits had 3.2-mm diameter tunnels drilled in the femur and tibia of both hind-limb stifle joints parallel to but without violation of the native anterior cruciate ligament (ACL). The animals were euthanized at 1, 2, 4, and 12 weeks postoperatively. Decalcified sections were made of the bone tunnels and new bone formation was computer quantified using histomorphometric methods at each time interval. Results: In this model, bone tunnel healing velocity was most rapid between 1 and 2 weeks after surgery. Both femoral and tibial interosseous tunnels showed substantial bone ingrowth (71% of bone tunnel volume) by 2 weeks postoperatively. The peripheral tunnel segment, that third of the tunnel furthest from the joint surface, healed rapidly and was 99% occluded with bone (99% confidence interval, 93.7% to 100%) at 2 weeks. Tunnel ingrowth was delayed and incomplete in the articular third of the tunnel, especially the femoral side. At 12 weeks, by volume, only 69.1% (99% confidence interval, 52.3% to 85.7%) of the interarticular third of the femoral tunnel was ingrown with new bone. Peripheral third bone tunnel healing was significantly greater than articular third tunnel healing at all time intervals; P <. 005 for the femoral and P <. 05 for the tibial tunnel. Conclusions: Interarticular bone tunnels heal from the outside in. At 12 weeks, bone healing was slower and incomplete in the articular segment of the tunnel, closest to the joint surface. The same biologic factors that impede intersubstance ACL healing may interfere with bone tunnel healing and be another cause of bone tunnel enlargement after ACL reconstruction.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 17, No 2 (February), 2001: pp 189–195     

Tendon healing in a bone tunnel. Part I: Biomechanical results after biodegradable interference fit fixation in a model of anterior cruciate ligament reconstruction in sheep

Purpose: Interference fit fixation of soft-tissue grafts has recently raised strong interest because it allows for anatomic graft fixation that may increase knee stability and graft isometry. Although clinical data show promising results, no data exist on how tendon healing progresses using this fixation. The purpose of the present study was to investigate anterior cruciate ligament (ACL) reconstruction biomechanically using direct tendon-to-bone interference fit fixation with biodegradable interference screws in a sheep model. Type of Study: Animal study. Methods: Thirty-five mature sheep underwent ACL reconstruction with an autologous Achilles tendon split graft. Grafts were directly fixed with poly-(D,L-lactide) interference screws. Animals were euthanized after 6, 9, 12, 24, and 52 weeks and standard biomechanical evaluations were performed. Results: All grafts at time zero failed by pullout from the bone tunnel, whereas grafts at 6 and 9 weeks failed intraligamentously at the screw insertion site. At 24 and 52 weeks, grafts failed by osteocartilaginous avulsion. At 24 weeks, interference screws were macroscopically degraded. At 6 and 9 weeks tensile stress was only 6.8% and 9.6%, respectively, of the graft tissue at time zero. At 52 weeks, tensile stress of the reconstruction equaled 63.8% and 47.3% of the Achilles tendon graft at time zero and the native ACL, respectively. A complete restitution of anterior-posterior drawer displacement was found at 52 weeks compared with the time-zero reconstruction. Conclusions: It was found that over the whole healing period the graft fixation proved not to be the weak link of the reconstruction and that direct interference fit fixation withstands loads without motion restriction in the present animal model. The weak link during the early healing stage was the graft at its tunnel entrance site, leading to a critical decrease in mechanical properties. This finding indicates that interference fit fixation of a soft-tissue graft may additionally alter the mechanical properties of the graft in the early remodeling stage because of a possible tissue compromise at the screw insertion site. Although mechanical properties of the graft tissue had not returned to normal at 1 year compared with those at time zero, knee stability had returned to normal at that time. There was no graft pullout after 24 weeks, indicating that screw degradation does not compromise graft fixation.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 18, No 2 (February), 2002: pp 113–123     

The use of platelets to affect functional healing of an anterior cruciate ligament (ACL) autograft in a caprine ACL reconstruction model

Many anterior cruciate ligament (ACL) reconstructions have increased laxity postoperatively. We hypothesized that enhancing an ACL graft with a collagen‐platelet composite (CPC) would improve knee laxity and graft structural properties. We also hypothesized the platelet concentration in the CPC would affect these parameters. Twelve goats underwent ACL reconstruction with autologous patellar tendon graft. In six goats, a collagen‐platelet composite was placed around the graft (CPC group). In the remaining six goats, the collagen scaffold only was used (COLL group). Three goats were excluded due to complications. After 6 weeks in vivo, anterior–posterior (AP) laxity and tensile properties of the ACL reconstructed knees were measured and normalized against the contralateral intact knee. At a knee flexion angle of 30°, the average increase in AP laxity was 40% less in the CPC group than the COLL group (p = 0.045). At 60°, the AP laxity was 30% less in the CPC group, a difference that was close to statistical significance (p = 0.080). No differences were found between treatment groups with respect to the structural properties (p > 0.30). However, there were significant correlations between serum platelet concentration and AP laxity (R² = 0.643; p = 0.009), maximum load (R² = 0.691; p = 0.006), and graft stiffness (R² = 0.840; p < 0.001). In conclusion, use of a CPC to enhance healing of an allograft ACL reconstruction inversely correlated with early sagittal plane laxity and the systemic platelet count was highly predictive of ACL reconstruction graft strength and stiffness at 6 weeks. These findings emphasize the importance of further research on delineating the effect of platelets in treating of ACL injuries. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 631–638, 2009     

保留与切除残迹前交叉韧带重建关节液渗入骨隧道的实验比较

目的探索前交叉韧带(ACL)重建术中保留与切除残迹对滑液渗入骨隧道的影响。方法将离体新鲜山羊膝关节20个,随机分为两组:保留残迹组和切除残迹组各10个关节,切断ACL,分别保留和完全切除ACL残迹,取跟腱为移植物,行ACL重建。于胫骨隧道做一骨窗显露移植腱,安放电极,向关节腔中注射10%的NaCl溶液20 ml,用电阻仪测量不同时间点(0、5、10、15、20 min)电阻值的变化。再向关节腔内注射碳素墨水20 ml,通过高清摄像头记录墨水渗出的时间。结果灌注后5、10、15 min,保留残迹组的电阻值明显高于切除残迹组,两组间差异有统计学意义(P<0.01)。保留残迹组的墨水渗出时间大于切除残迹组,两组间差异有统计学意义(P<0.01)。结论保留残迹ACL重建可以明显减少或延缓关节液渗入骨隧道。